LTP induction initiated by Ca2+ influx through the NMDA receptor leads to CaMKII activation and translocation to the postsynaptic density, where it is localized by binding to the NMDAR. The NMDAR has a unique role as the only known activity-dependent binding partner for CaMKII within the PSD. This proposal will address the roles of 2 NMDAR CaMKII binding domains, NR1-CO and NR2B-C, in the specificity of synaptic CaMKII signaling. Aim 1 characterizes the roles of NR1-CO/NR2B-C in synaptic CaMKII targeting and substrate phosphorylation. Aim 2 characterizes the roles of NR1-CO/NR2B-C in the structural organization of the synapse. The proposed experimental design utilizes NR1-CO and NR2B-C knock-in mice as a powerful strategy in combination with biochemical (immunoprecipitation and subcellular fractionation) and cellular (immunofluorescence in hippocampal cultures) techniques. The findings of this proposal will provide insight into the molecular mechanisms of LTP, as well as lead to new directions for targeted therapies in stroke, as similar phenomena occur following LTP induction and cerebral ischemia.